高效液相色谱串联质谱电喷雾检测法快速测定人尿中帕洛诺司琼的浓度

Rapid determination of palonosetron in urine by LC-ESI-MS/MS

目的:建立快速测定人尿中帕洛诺司琼浓度的高效液相色谱串联质谱电喷雾检测法(LC-ESI-MS/MS)。方法:以AgilentTCC18柱(4.6mm×150mm,5μm)为色谱柱;流动相为乙腈-0.04mol.L-1甲酸铵水溶液(含0.04%甲酸)(69∶31),流速0.6mL·min-1;柱温25℃;醋酸乙酯-二氯甲烷(4∶1)为提取剂。样品经电喷雾离子源正离子化后,样品通过三重四级杆串联质谱仪,采用选择反应监测(SRM)对帕洛诺司琼(m/z297.2→82.2)和内标地西泮(m/z285.1→154.0)测定。结果:帕洛诺司琼高(80μg.L-1)、中(50μg.L-1)、低(5μg.L-1)3个浓度的平均回收率RSD均小于15%;线性范围为2.5～100μg.L-1,回归方程为F=27.135ρ-0.0582,r=0.9999(n=6),权重系数为1/ρ,分析方法的最低定量限为2.5μg.L-1。结论:该方法灵敏、准确、简单、快速,可用于帕洛诺司琼临床尿药浓度监测和药动学研究。

OBJECTIVE To set up a LC-ESI-MS/MS method for palonosetron assay in urine. METHODS Palonosetron was extracted with ethyl acetate dichloromethane. The residues were analyzed with a LOESI MS/MS system （Agilent TC C18 column,4. 6 mm× 150 mm, 5 μm） with the mobile phase consisted of acetonitrile with 0. 04 mmol, L^-1 ammonium formate （0. 04% formic acid ） = 69：31 （V： V）. The flow rate was 0. 6 mL·min ^-1. A Agilent 6410 triple quad mass spectrometer sys tern equipped with an electrospray ionization ion trap source was used as the detector and operated in the positive ion mode. Se lected reaction monitoring（SRM） using the precursor to produce combinations of m/z 297. 2→82. 2 and m/z 285. 1→154. 0 was performed to detect palonosetron and the internal standard, respectively. RESULTS The average recoveries for palonosetron were 69.00% ,69.75% ,71.61% ,respectively. The within-day （n = 5） and between day （n = 3） precision of variation was less than 15 %. The calibration curves for palonosetron had good linearity, r = 0. 999 9（n = 6）,over the range of 2.5 - 100 μg·L^-1. The limits of quantitation for palonosetron was 2. 5 μg·L^-1. CONCLUSION The method provides a sensitive,accurate, precise and reliable analytical procedure for clinical monitoring of palonosetron in urine and for its phamacokinetic studies.